重組人Dickkopf1蛋白的克
發(fā)布時間:2018-02-27 21:03
本文關(guān)鍵詞: dickkopf1 Dkk1蛋白 黑素細(xì)胞 黑素 酪氨酸酶 出處:《第四軍醫(yī)大學(xué)》2006年碩士論文 論文類型:學(xué)位論文
【摘要】:黑素是皮膚成為褐色的主要色素。黑素是酪氨酸在酪氨酸酶作用下產(chǎn)生的,在黑素細(xì)胞的黑素小體中生成,黑素小體從一個黑素細(xì)胞被轉(zhuǎn)運(yùn)至一群角質(zhì)形成細(xì)胞。真、表皮內(nèi)黑素細(xì)胞增生及黑素生成功能活躍是色素增多性疾病的一個主要原因。色素增多性疾病在皮膚科疾病中非常常見,最常見的是雀斑、黃褐斑及炎癥后色素沉著,,發(fā)生在面部、手背等暴露部位顯著者嚴(yán)重影響患者的容貌,影響患者的生活質(zhì)量。根據(jù)色素增多性疾病的發(fā)病機(jī)制,防治措施主要從抑制黑素細(xì)胞增殖及活性、防止黑素生成及加速黑素降解等方面入手。目前針對色素增多性疾病防治的各種方法的療效、復(fù)發(fā)問題以及可能出現(xiàn)的不良反應(yīng),一直是皮膚科臨床醫(yī)師所面臨的棘手問題。 人掌跖部位皮膚的黑素含量比非掌跖部位低。近來研究發(fā)現(xiàn),掌跖部位皮膚黑素細(xì)胞的密度是非掌跖部位皮膚的1/5,黑素小體、黑素相關(guān)蛋白含量也相應(yīng)的顯著降低,其原因為掌跖部位皮膚中的成纖維細(xì)胞分泌的Dickkopf1蛋白能夠通過抑制黑素細(xì)胞Wnt信號轉(zhuǎn)導(dǎo)通路的活化進(jìn)而抑制黑素細(xì)胞的增殖、發(fā)育、分化及黑素生成能力。 本課題通過反轉(zhuǎn)錄PCR自人掌跖部位皮膚成纖維細(xì)胞中獲得了編碼Dickkop1的32—266位氨基酸的基因,克隆入PQE30質(zhì)粒,構(gòu)建了PQE30-dkk1原核表達(dá)質(zhì)粒,經(jīng)雙酶切鑒定及序列分析證實所獲序列完全正
[Abstract]:Melanin is the primary brown pigment of the skin. Melanin is produced by tyrosine acting on tyrosinase, formed in the melanoma body of melanocytes, which is transported from a melanocyte to a group of keratinocytes. Proliferation of melanocytes and active melanogenesis in the epidermis are one of the main causes of hyperpigmentation. Hyperpigmentation is very common in dermatology, and the most common diseases are freckles, chloasma and postinflammatory pigmentation. Those with prominent exposure sites such as the face and the back of the hand seriously affect the appearance of the patient and the quality of life of the patient. According to the pathogenesis of pigmentation disease, the prevention and treatment measures mainly include inhibiting the proliferation and activity of melanocytes. Prevention of melanogenesis and acceleration of melanin degradation, etc. At present, the curative effect, recurrence and possible adverse reactions of various methods for the prevention and treatment of hyperpigmented diseases have been the thorny problems faced by dermatologists. The melanin content of human palmoplantar skin is lower than that of non-palmoplantar skin. Recently, it has been found that the density of melanocytes in palmar plantar skin is 1 / 5 of that in non-palmar metatarsal skin, and melanosome and melanin related protein content also decrease significantly. The reason is that the Dickkopf1 protein secreted by fibroblasts in palmar plantar skin can inhibit the proliferation, development, differentiation and melanogenesis of melanocytes by inhibiting the activation of Wnt signal transduction pathway of melanocytes. In this study, the gene encoding 32-266 amino acids of Dickkop1 was obtained from human palmoplantar skin fibroblasts by reverse transcription PCR and cloned into PQE30 plasmid. The prokaryotic expression plasmid of PQE30-dkk1 was constructed and confirmed by double enzyme digestion and sequence analysis.
【學(xué)位授予單位】:第四軍醫(yī)大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2006
【分類號】:R346
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